Zhenwu decoction ameliorates adriamycin-induced nephrotic syndrome in rats via dual modulation of AVP-V2R-AQP2 and RAAS-MR-AQP3 pathways

真武汤通过双重调节 AVP-V2R-AQP2 和 RAAS-MR-AQP3 通路改善阿霉素诱导的大鼠肾病综合征

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Abstract

The core bioactive components of Zhenwu Decoction (ZWD)-utilized for managing kidney diseases-and the associated anti-nephrotic syndrome (NS) edema mechanism remain incompletely characterized. We assessed the potential mechanism of ZWD against adriamycin (ADR)-induced NS. Sprague-Dawley rats (n = 6/group) received intravenous adriamycin (6 mg/kg) to establish NS models and were administered ZWD (4.2, 8.4, 16.8 g/kg/day) and prednisone (5 mg/kg/day) for 4 weeks. Renal function was assessed using the serum biomarkers: blood urea nitrogen (BUN), serum total protein (TP), creatinine (Scr), albumin (ALB), triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). H&E and Masson's trichrome staining were used to assess the pathological alterations and renal fibrosis status. Enzyme-linked immunosorbent assay was used to detect the serum levels of arginine vasopressin (AVP), renin (Ren), angiotensin II (AngII), and aldosterone (ALD). Immunohistochemistry, Western blot, and RT-qPCR were performed to examine the levels of relevant intracellular renal signaling molecules. ZWD treatment significantly reduced proteinuria by 58% in NS rats, along with marked improvements in renal pathology and fibrosis. ZWD also ameliorated serum levels of TP and LDL-C, and lowered the concentrations of AVP, Renin, AngII, and ALD. The expression of type 2 vasopressin receptor (V2R), mineralocorticoid receptor (MR), aquaporin 1 (AQP1), AQP2, and AQP3 was remarkably decreased at the protein and mRNA levels. Three components (benzoylpaeoniflorin, 6-gingerol, and paeoniflorin) of ZWD exhibited synergistic effects on AQP1, AQP2, AQP3, MR, and V2R. Our findings suggest that ZWD alleviates NS through renal repair and dual suppression of the AVP-V2R-AQP2 and RAAS-MR-AQP3 pathways.

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